title: Clonorchis sinensis enolase: identification and biochemical characterization of a glycolytic enzyme from excretory/secretory products.
abstract: Enolase plays a key role in energy metabolism and development of most organisms. We isolated a gene encoding enolase from Clonorchis sinensis (C. sinensis) adult cDNA library and expressed the recombinant protein in Escherichia coli. C. sinensis enolase (Csenolase) was identified as both an excretory/secretory product and a tegumental component of C. sinensis by western blot analysis. The transcriptional level of Csenolase was examined at adult worm, metacercaria, cercaria and egg of C. sinensis, and results showed that Csenolase is transcribed at the four life stages of C. sinensis while showing a significant higher expression level at the stage of adult worm. Immunohistochemical localization indicated that Csenolase was specifically deposited on the tegument of adult worm and cyst wall of metacercaria. Ligand blot assay revealed a specific characteristic of dose-dependent plasminogen-binding activity of Csenolase and kinetic parameters were explored using 2-phospho-D-glycerate (2-PGA) as the primary substrate by monitoring the conversion of nicotinamide-adenine dinucleotide (NADH) into nicotinamide adenine dinucleotide (NAD). In addition, Csenolase exhibited active enzyme activity in catalytic reactions while the anti-Csenolase serum inhibited the enzyme activity. In vitro incubation experiments revealed that Csenolase might play key roles in the growth of the parasites. In conclusion, Csenolase is an important glycolytic enzyme required for the development of C. sinensis, and may be a potential vaccine candidate and drug target against C. sinensis infection.
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, PR China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wang
authors.forename: Wenjun
authors.initials: W
authors.lastname: Chen
authors.forename: Fengyu
authors.initials: F
authors.lastname: Hu
authors.forename: Chuanhuan
authors.initials: C
authors.lastname: Deng
authors.forename: Chenhui
authors.initials: C
authors.lastname: Zhou
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Lv
authors.forename: Yongxiu
authors.initials: Y
authors.lastname: Fan
authors.forename: Jingtao
authors.initials: J
authors.lastname: Men
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.forename: Jiufeng
authors.initials: J
authors.lastname: Sun
authors.forename: Dong
authors.initials: D
authors.lastname: Hu
authors.forename: Jingfang
authors.initials: J
authors.lastname: Chen
authors.forename: Yabo
authors.initials: Y
authors.lastname: Yang
authors.forename: Chi
authors.initials: C
authors.lastname: Liang
authors.forename: Huanqin
authors.initials: H
authors.lastname: Zheng
authors.forename: Xuchu
authors.initials: X
authors.lastname: Hu
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.forename: Zhongdao
authors.initials: Z
authors.lastname: Wu
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000201:enzymology* / Q000235:genetics*; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D020869:Gene Expression Profiling; D007150:Immunohistochemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000378:metabolism*; D010958:Plasminogen / Q000378:metabolism; D011485:Protein Binding; D017422:Sequence Analysis, DNA
chemical_list: D018509:DNA, Helminth; D010958:Plasminogen; D010751:Phosphopyruvate Hydratase